Effect of Ball-Milling Treatment Combined with Glycosylation on the Structure and Functional Properties of Litopenaeus vannamei Protein.

Litopenaeus vannamei protein (LVP) is a high-quality protein. However, its functional properties do not fully meet the needs of food processing. In this study, LVP-xylose conjugates were prepared by conventional wet heat method (GLVP) and ball-milling-assisted wet heat method (GBLVP), respectively. The changes in structure and functional properties of the glycosylated LVP were explored. The findings revealed that ball-milling pretreatment increased the grafting degree to 35.21%. GBLVP had a sparser surface structure and lower particle size than GLVP. FTIR spectra showed that xylose was grafted onto LVP successfully and GBLVP had the lowest α-helix content. Compared with GLVP, GBLVP had a decrease in intrinsic fluorescence intensity and surface hydrophobicity, and an increase in UV absorption intensity. Moreover, GBLVP had higher foaming capacity, solubility and water-holding capacity, and lower allergenicity than GLVP. However, ball-milling pretreatment had a negative impact on the vitro digestibility and oil-holding capacity of GBLVP. In conclusion, ball-milling-assisted treatment of glycosylation could effectively improve the functional properties of LVP, benefiting the broader application of LVP in the food industry.

Enhancing the endo-activity of the thermophilic chitinase to yield chitooligosaccharides with high degrees of polymerization.

Thermophilic endo-chitinases are essential for production of highly polymerized chitooligosaccharides, which are advantageous for plant immunity, animal nutrition and health. However, thermophilic endo-chitinases are scarce and the transformation from exo- to endo-activity of chitinases is still a challenging problem. In this study, to enhance the endo-activity of the thermophilic chitinase Chi304, we proposed two approaches for rational design based on comprehensive structural and evolutionary analyses. Four effective single-point mutants were identified among 28 designed mutations. The ratio of (GlcNAc)3 to (GlcNAc)2 quantity (DP3/2) in the hydrolysates of the four single-point mutants undertaking colloidal chitin degradation were 1.89, 1.65, 1.24, and 1.38 times that of Chi304, respectively. When combining to double-point mutants, the DP3/2 proportions produced by F79A/W140R, F79A/M264L, F79A/W272R, and M264L/W272R were 2.06, 1.67, 1.82, and 1.86 times that of Chi304 and all four double-point mutants exhibited enhanced endo-activity. When applied to produce chitooligosaccharides (DP ≥ 3), F79A/W140R accumulated the most (GlcNAc)4, while M264L/W272R was the best to produce (GlcNAc)3, which was 2.28 times that of Chi304. The two mutants had exposed shallower substrate-binding pockets and stronger binding abilities to shape the substrate. Overall, this research offers a practical approach to altering the cutting pattern of a chitinase to generate functional chitooligosaccharides.

Effects of different high temperature-pressure processing times on the sensory quality, nutrition and allergenicity of ready-to-eat clam meat.

Investigating technologies to control the allergenicity of seafood is particularly important to safeguard consumer health, but there is currently a dearth of research focused on reducing the allergenicity of clam meat. This study aimed to investigate the effects of high temperature-pressure (HTP) processing times (121 °C, 0.14 MPa; 5, 10, 15, 20 min) on the sensory quality, nutrition, and allergenicity of ready-to-eat clam meat. With the extension of HTP time, the hardness of clam meat gradually decreased, the chewiness decreased initially and then increased, and the meat became tender. HTP processing endowed clam meat with abundant esters and aldehydes. Among all the processing groups, the umami and saltiness were better at 15 min, correlating with the highest overall acceptability. Ready-to-eat clam meat contained high-protein nutritional value. Compared with raw clam meat, the tropomyosin allergenicity of clam meat treated with HTP for 15 and 20 min was significantly reduced by 51.9 % and 56.5 %, respectively (P < 0.05). However, there was no significant difference between these two groups. Appropriate HTP processing time might be an efficient condition to reduce the tropomyosin allergenicity of ready-to-eat clam meat and improve its quality, particularly for the time of 15 min. The results of this study could provide a reliable theoretical basis for the development of hypoallergenic clam foods.

Optimization of Extraction and Refining Parameters of Oil from Dotted Gizzard Shad (Konosirus punctatus).

To address the challenges associated with resource inefficiency, low extraction rates, environmental concerns, and high energy consumption in traditional fish oil production from dotted gizzard shad (Konosirus punctatus), a novel approach is needed. This study aimed to develop and evaluate two innovative methods for fish oil extraction and refinement, focusing on their effects on fish oil quality, fatty acid profile, and volatile compound composition throughout the respective processes. The findings of the study revealed that the ethanol-assisted enzymatic extraction method surpassed the conventional enzymatic approach in extraction efficiency, achieving an optimal extraction rate of 74.94% ± 0.45% under optimized process conditions. Moreover, the ethanol-NaOH one-step degumming and deacidification method proved effective in simultaneously removing phospholipids and free fatty acids. Under optimal conditions, a notable reduction in phospholipid content in dotted gizzard shad oil, from 6.80 ± 0.01 mg/g to 1.18 ± 0.01 mg/g, and a substantial decrease in acid value, from 3.31 mg/g to 0.31 mg/g, were observed. In summary, the study analyzed the physicochemical properties, fatty acid composition, and volatile components of fish oil before and after refinement. The refining process was found to preserve the fatty acid composition while efficiently eliminating hydroperoxides and reducing unpleasant odors in the crude oil.

Integrated ultra-high pressure and salt addition to improve the in vitro digestibility of myofibrillar proteins from scallop mantle (Patinopecten yessoensis).

Myofibrillar protein (MP) is susceptible to the effect of ionic strength and ultra-high pressure (UHP) treatment, respectively. However, the impact of UHP combined with ionic strength on the structure and in vitro digestibility of MP from scallop mantle (Patinopecten yessoensis) is not yet clear. Therefore, it is particularly important to analyze the structural properties and enhance the in vitro digestibility of MP by NaCl and UHP treatment. The findings demonstrated that as ionic strength increased, the α-helix and ß-sheet gradually transformed into ß-turn and random coil. The decrease of endogenous fluorescence intensity indicated the formation of a more stable tertiary structure. Additionally, the exposure of internal sulfhydryl groups increased the amount of total sulfhydryl content, and reactive sulfhydryl groups gradually transformed into disulfide bonds. Moreover, it reduces aggregation through increased solubility, decreased turbidity, particle sizes, and a relatively dense and uniform microstructure. When MP from the scallop mantle was treated with 0.5 mol/L ionic strength and 200 MPa UHP treatment, it had the highest solubility (90.75 ± 0.13%) and the lowest turbidity (0.41 ± 0.03). The scallop mantle MP with NaCl of 0.3 mol/L and UHP treatment had optimal in vitro digestibility (95.14 ± 2.01%). The findings may offer a fresh perspectives for developing functional foods for patients with dyspepsia and a theoretical foundation for the comprehensive utilization of scallop mantle by-products with low concentrations of NaCl.

Improvement in the gelling properties of myofibrillar protein from the razor clam (Sinonovacula constricta) through phosphorylation and structural characterization of the modified protein.

This study investigated the modification of myofibrillar protein (MP) from the razor clam through phosphorylation by using various phosphate salts, namely, sodium tripolyphosphate (STPP), sodium trimetaphosphate (STMP), sodium polyphosphate (STTP) and sodium pyrophosphate (TSPP), and their mechanisms of action for functional and gelling properties. Fourier transform infrared spectrometry (FTIR) showed that MP introduced phosphate groups during phosphorylation; these phosphates changed the secondary structure. Moreover, MP after phosphorylation led to an increase in solubility, which was more evident in the case of TSPP phosphorylation, leading to the improvement of gel properties. Therefore, TSPP was the phosphate with the best gel properties in the modification of MP, showing the highest phosphorus content, which resulted in better gelling properties owing to its relatively shorter chains. These results showed that phosphate was able to improve protein cross-linking through ion interactions and electrostatic interactions, which ultimately improved the gelling properties of the razor clam protein.

Effects of ultrasound-assisted high temperature-pressure treatment on the structure and allergenicity of tropomyosin from clam (Mactra veneriformis).

Tropomyosin (TM) is the major allergen in clams. This study aimed to evaluate the effects of ultrasound-assisted high temperature-pressure treatment on the structure and allergenicity of TM from clams. The results showed that the combined treatment significantly affected the structure of TM-converting the α-helix to ß-sheet and random coil, and decreasing the sulfhydryl group content, surface hydrophobicity, and particle size. These structural changes caused the unfolding of the protein, disrupting and modifying the allergenic epitopes. The significant reduction in the allergenicity of TM was approximately 68.1% when treated with combined processing (P < 0.05). Notably, an increase in the content of the relevant amino acids and a smaller particle size accelerated the penetration of the enzyme into the protein matrix, resulting in strengthening the gastrointestinal digestibility of TM. These results prove that ultrasound-assisted high temperature-pressure treatment has great potential in reducing allergenicity, benefiting the development of hypoallergenic clam products.

Exploring the Fungal Community and Its Correlation with the Physicochemical Properties of Chinese Traditional Fermented Fish (Suanyu).

Suanyu is a traditional natural fermented fish product from Southwest China that contains very complex microflora. The main purpose of this study was to explore the fungal community and its relationship with the physicochemical properties of Suanyu. The fungal community structure of Suanyu from the main provinces (Guizhou and Hunan) was studied via high-throughput sequencing. The correlation between dominant fungi and physicochemical characteristics was analyzed via Spearman's correlation coefficient. The results showed that the pH value, total volatile base nitrogen content, and thiobarbituric acid reactive substance content ranges of Suanyu samples were 4.30-5.50, 17.11-94.70 mg/100 g, and 0.61 to 3.62 mg/kg, respectively. The average contents of total volatile base nitrogen, thiobarbituric acid reactive substance, and total BAs in Suanyu from Guizhou were lower than those from Hunan. The main BAs were phenethylamine, putrescine, cadaverine, histamine, and tyramine. Ascomycota was the dominant fungal phylum, and Kodamaea, Debaryomyces, Wallemia, Zygosaccharomyces, and unclassified Dipodascaceae were the dominant fungal genera in different samples. Moreover, high abundance levels of Kodamaea and Zygosaccharomyces were found in Suanyu from Guizhou. According to the correlation analysis, Kodamaea and Zygosaccharomyces were negatively correlated with TBARS (R2 = -0.43, -0.51) and TVBN (R2 = -0.37, -0.29), and unclassified Dipodascaceae was significant negatively correlated with tyramine (R2 = -0.56). This study expands the understanding of the fungal community and the fermentation characteristics of the dominant fungi in Suanyu.

Whole-genome sequencing and functional analysis of a novel chitin-degrading strain Rhodococcus sp. 11-3.

Chitin is the second most abundant polysaccharide in nature. Therefore, how to utilize the resource is an important issue. Rhodococcus sp. 11-3 is a strain with high chitin deacetylase (CDA) activity. In the present study, we used a combined Illumina and PacBio sequencing strategy to assemble the whole genome sequence of this strain. The genome of Rhodococcus sp. 11-3 was 5,627,661 bp in size and contained 5983 coding genes, of which 5983, 4040, 4648, 4914, 4174, 2350, and 173 genes were annotated in the Non-Redundant Protein Database (NR), Swiss-Prot, Pfam, Clusters of Orthologous Groups of proteins (COG), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Carbohydrate-active enzymes (CAZymes) databases, respectively. The genome was annotated to a chitin deacetylase gene (RhoCDA) and a chitinase gene (RhoChi). They were not very similar to the previously reported chitin deacetylase and chitinase. This made it possible to investigate the genes associated with chitin degradation and would provide an important reference for subsequent gene cloning, functional research, development and application. Therefore, the Rhodococcus sp. 11-3 strain has great potential in the development of chitin resources.

Variations in the physicochemical properties and bacterial community composition during fermentation of low-salt shrimp paste.

The main purpose of the present study was to investigate the influence of two fermentation temperatures (10 °C and 20 °C) on the physicochemical properties and bacterial community composition of the low-salt shrimp paste. The physicochemical properties pH values, total volatile basic nitrogen (TVBN), amino acid nitrogen (AAN) and biogenic amines (BAs) of low-salt shrimp paste were analyzed and compared. The bacterial community composition was analyzed by high-throughput sequencing technology. Results indicate that the pH value fluctuated between 8.50 and 7.95, and the AAN, TVBN and total BAs levels of low-salt shrimp paste increased gradually after fermentation started, but the AAN content fluctuated steadily after 12 w. The contents of TVBN and total BAs in shrimp paste fermented at 10 °C were lower than those at 20 °C. Thus, the appropriate fermentation temperature and period were 10 °C and 12 w, respectively. Carnobacteriaceae and Staphylococaceae were the predominant family for the fermented shrimp paste. At the genus level, Alkalibacterium was predominantly present at 8 wk (96.36%), while Staphylococcus became the predominant one at 20 wk (58.71%). Correlation analysis shows that Alkalibacterium was positively associated with tyramine (R2 = 0.59), and Staphylococcus and Tetragenococcus were positively associated with AAN and TVBN. These findings are used to facilitate the quality control and improvement of low-salt shrimp paste production process in the future.

A novel thermophilic chitinase directly mined from the marine metagenome using the deep learning tool Preoptem.

Chitin is abundant in nature and its degradation products are highly valuable for numerous applications. Thermophilic chitinases are increasingly appreciated for their capacity to biodegrade chitin at high temperatures and prolonged enzyme stability. Here, using deep learning approaches, we developed a prediction tool, Preoptem, to screen thermophilic proteins. A novel thermophilic chitinase, Chi304, was mined directly from the marine metagenome. Chi304 showed maximum activity at 85 â„ƒ, its Tm reached 89.65 ± 0.22℃, and exhibited excellent thermal stability at 80 and 90 °C. Chi304 had both endo- and exo-chitinase activities, and the (GlcNAc)2 was the main hydrolysis product of chitin-related substrates. The product yields of colloidal chitin degradation reached 97% within 80 min, and 20% over 4 days of reaction with crude chitin powder. This study thus provides a method to mine the novel thermophilic chitinase for efficient chitin biodegradation.

Comprehensive Evaluation of Volatile and Nonvolatile Compounds in Oyster Cuts of Roasted Lamb at Different Processing Stages Using Traditional Nang Roasting.

Nang roasting is a traditional lamb processing method in Xinjiang (China) with a history of thousands of years. This study comprehensively evaluated the volatile and nonvolatile compounds of oyster cuts of roasted lamb at different processing stages of Nang roasting using gas chromatography mass spectrometry and amino acid automatic analyzer, respectively. Results indicated that aldehydes were the dominant profiles of volatile compounds, and hexanal, nonanal, octanal, (E)-2-nonenal, (E, E)-2,4-decadienal, (E, E)-2,4-nonadienal and 1-octen-3-ol were the key volatile compounds or aroma contributors to roasted oyster cuts. Isoamylol and 3-hydroxy-2-butanone could differentiate fresh and marinated oyster cuts from roasted ones; (E)-2-nonenal, (E, E)-2,4-decadienal, 1-octen-3-ol, hexanal, octanal, nonanal and (E, E)-2,4-nonadienal could differentiate Nang roasted oyster cuts of 60 min from those of 15, 30 and 45 min. Umami amino acids and sweet amino acids are the dominant profiles of nonvolatile compounds; glutamic acid, alanine and 5'-IMP were the key free amino acids or taste contributors to roasted oyster cuts. Glutamic acid, alanine and 5'-IMP could differentiate fresh and marinated oyster cuts from roasted samples. This work provided theoretical support for the control of flavor attributes of roasted lamb with traditional Nang roasting.

Identification of a chitosanase from the marine metagenome and its molecular improvement based on evolution data.

Chitooligosaccharides have important application value in the fields of food and agriculture. Chitosanase can degrade chitosan to obtain chitooligosaccharides. The marine metagenome contains many genes related to the degradation of chitosan. However, it is difficult to mine valuable genes from large gene resources. This study proposes a method to screen chitosanases directly from the marine metagenome. Chitosanase gene chis1754 was identified from the metagenome and heterologously expressed in Escherichia coli. The optimal temperature and pH of CHIS1754 were 55 °C and 5.5, respectively. A mutant, CHIS1754T, with 15 single point mutations designed based on molecular evolution data was also expressed in E. coli. The results indicated that the thermal stability of CHIS1754T was significantly improved, as the Tm showed an increase of ~ 7.63 °C. Additionally, the kcat/Km of CHIS1754T was 4.8-fold higher than that of the wild type. This research provides new theories and foundations for the excavation, modification, and industrial application of chitosanases. KEY POINTS: A chitosanase gene, chis1754, was firstly identified from marine metagenome. A multi-site mutant was designed to improve enzyme stability and activity. The kcat/Kmof the designed mutant was 4.8-fold higher than that of the wild type.

Biological degradation of aflatoxin M1 by Bacillus pumilus E-1-1-1.

Aflatoxin M1 (AFM1 ) is a potent mycotoxin which causes serious health concerns in developing countries, where it is mainly found in milk, meat, and other foods. Biological detoxification is a promising method for eliminating AFM1 . The aim of this work was to search for AFM1 -degrading bacterial strains from animal waste, soil, and activated sludge. High-performance liquid chromatography and Fourier-transform infrared spectroscopy were used to analyze the AFM1 degradation products. A strain designated E-1-1-1 was obtained from African elephants feces, with the degradation ratio of AFM1 reaching 89.55% in 12 hr. Based on morphology, physiological and biochemical tests, and 16S rRNA gene sequence analysis, strain E-1-1-1 was identified as Bacillus pumilus. The culture supernatant of B. pumilus E-1-1-1 degraded AFM1 effectively, whereas the cells and cell extracts of B. pumilus E-1-1-1 were far less effective. Carbon and nitrogen sources had highly significant effects on the degradation of AFM1 by B. pumilus E-1-1-1. The AFM1 -degrading strain, B. pumilus E1-1-1, could have great potential in industrial applications.

Metabolic profiling of Staphylococcus aureus cultivated under aerobic and anaerobic conditions with (1)H NMR-based nontargeted analysis.

Staphylococcus aureus is a major pathogen in the medical area and food-producing sector. Detailed analyses of its basic cell physiology will help comprehensively understand this pathogen, which will be useful for developing novel diagnostic and treatment tools. Oxygen is one of the most crucial growth-limiting factors for S. aureus. In this study, to characterize and distinguish metabolic profiles of S. aureus cultivated under aerobic and anaerobic conditions, nontargeted analyses of both types of cultures were carried out using (1)H nuclear magnetic resonance spectroscopy. Fifty compounds were identified by Chenomx software. Characteristics of metabolic profiles were achieved by using principal components analysis. During aerobic growth, S. aureus mainly consumed glucose, alanine, arginine, glycine, isoleucine, leucine, phenylalanine, and acetate. Meanwhile, it accumulated 17 metabolites, mainly 2-oxoglutarate, isobutyrate, isovalerate, succinate, and ethanol. Under anaerobic condition, S. aureus mainly consumed glucose, arginine, and threonine. Meanwhile, it accumulated 13 metabolites, mainly ethanol, lactate, and ornithine. The representative metabolites that could most significantly differentiate metabolic profiles of S. aureus were isobutyrate, isovalerate, and succinate in aerobic cultivation; and lactate, ethanol, and ornithine in anaerobic cultivation. Among these metabolites, isobutyrate and ornithine were present only in aerobic and anaerobic culture, respectively.

Efficacy of acidic and basic electrolyzed water in eradicating Staphylococcus aureus biofilm.

Staphylococcus aureus is a major pathogen. It can form biofilm on the surfaces of medical devices and food equipment, which makes it more difficult to eradicate. To develop a novel method to eradicate S. aureus biofilm, the effects of electrolyzed water on removing and killing S. aureus biofilm were investigated in this study. By using a biofilm biomass assay with safranin staining and visualization of biofilm architecture with scanning electron microscopy, it was shown that basic electrolyzed water (BEW) could effectively remove established biofilm. The pH of electrolyzed water affected removal efficacy. Using a biofilm viability assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide staining, acidic electrolyzed water (AEW) efficiently killed biofilm-imbedded S. aureus. The available chlorine in AEW may be a main contributing factor for bactericidal activity. Additionally, BEW had a removal efficacy for S. aureus biofilm equivalent to 2% NaOH, and AEW had a bactericidal capability for S. aureus in biofilm equivalent to 2% HCl. These data suggested that AEW and BEW could be applied as a bactericide and removing agent for S. aureus in biofilm, respectively.